Establishment and optimization of Agrobacterium mediated genetic transformation system for Magnaporthe oryzae

doi:10.14088/j.cnki.issn0439-8114.2025.03.029

Abstract

Abstract: ： Magnaporthe oryzae was used as the experimental material. The GFP gene was obtained through PCR, and a pBarg-GFP-BAR-Amp recombinant vector was constructed. The Magnaporthe oryzae strain containing the GFP gene with stable inheritance was obtained using Agrobacterium-mediated transformation. The transformation conditions were optimized using single factor and response surface methodology. The optimal transformation conditions for the Magnaporthe oryzae genetic transformation system were an OD_{600 nm} of 0.38 （Agrobacterium concentration）, a co-culture temperature of 28.2 ℃, a co-culture time of 2.12 days, and a transformation efficiency of 280.00×10^-5. PCR identification and fluorescence microscopy observation revealed that the GFP gene of the transformant Magnaporthe oryzae could be expressed normally, with no significant difference in pathogenicity compared to the wild-type Magnaporthe oryzae.

Key words: Magnaporthe oryzae, Agrobacterium mediated, genetic transformation, GFP gene, establish, optimization

CLC Number:

S432.4

LI Shuang, ZHANG Shu-mei, TIAN Yuan, PAN Yu, LIU Wei, YAN Geng-xuan. Establishment and optimization of Agrobacterium mediated genetic transformation system for Magnaporthe oryzae[J]. HUBEI AGRICULTURAL SCIENCES, 2025, 64(3): 182-189.

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